The production and utility of monoclonal antibodies to rat apolipoprotein B lipoproteins.

Murine hybridomas were prepared by fusion of azaguanine-resistant plasmacytoma cells and spleen cells from BALB/c mice immunized with intact rat lipoproteins (Lp). Seventeen stable hybridomas from 5 fusions producing antibody against rat apolipoprotein B (apo B)-containing Lp were prepared. The distribution of antigenic determinants of rat apo B variants defined by these antibodies was similar to that found for human apo B. Thirteen mAbs were reactive to epitopes found on both the higher (apo BH) and lower (apo BL) molecular weight forms of rat apo B; two were reactive to apo BH-specific epitopes and two were reactive to apolipoprotein E. No antibodies reactive to unique apo BL epitopes were found. One anti-rat apo B mAb (1.34.3) of high affinity and avidity was reactive to denatured and intact Lp-apo B. Reactivity towards Lp-apo B was independent of variant composition and density class of Lp. The mAb was subsequently used to develop a radioimmunoassay (RIA) which was sensitive down to 5 ng apo B. The assay was used successfully to measure total nascent Lp-apo B secreted by hepatocytes in culture.